Guo Yuehua, Liu Shihua, Yang Huili, Wang Po, Feng Qiumei
Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong, 226001, China.
School of Chemistry and Materials Science, Jiangsu Normal University, Xuzhou, 221116, China.
Anal Chim Acta. 2021 Feb 1;1144:68-75. doi: 10.1016/j.aca.2020.12.004. Epub 2020 Dec 7.
The sensitive detection of telomerase activity is of great significance for the early diagnosis and treatment of cancer. Here, an innovative electrochemiluminescence resonance energy transfer (ECL-RET) sensor was explored to reliably detect telomerase activity based on proximity binding-triggered multipedal DNA walker. In this system, CdS quantum dots (CdS QDs) and silver nanoclusters (Ag NCs) were applied as ECL donor and acceptor, respectively. By ingeniously introducing a repetitive bases sequence (TTAGGG) along the telomerase primer, multiple same DNA "legs" were formed, leading to the activation of proximity binding-triggered multipedal DNA walker. Unlike the traditional unipedal DNA walker, one walking step of multipedal DNA walker concurrently initiated the responsivity of multiple signals, resulting in the shortening of the walking time and improvement of the signal amplification efficiency. Thus, under optimal conditions, the designed ECL-RET sensor exhibited a wide dynamic correlation of HeLa cells' telomerase activity from 1 × 10 to 1 × 10 cells/mL and a low detection limit of 16 cells/mL. Moreover, this sensor realized the general and reliable analysis of telomerase activity in different cell lines. Due to the outstanding application potential in real samples, it is believed that the ECL-RET sensing system provides a new approach for the application of telomerase activity assays in cancer diagnostics.
端粒酶活性的灵敏检测对于癌症的早期诊断和治疗具有重要意义。在此,我们探索了一种创新的电化学发光共振能量转移(ECL-RET)传感器,用于基于邻近结合触发的多足DNA步行器可靠地检测端粒酶活性。在该系统中,硫化镉量子点(CdS QDs)和银纳米簇(Ag NCs)分别用作ECL供体和受体。通过沿着端粒酶引物巧妙地引入重复碱基序列(TTAGGG),形成了多个相同的DNA“腿”,从而激活了邻近结合触发的多足DNA步行器。与传统的单足DNA步行器不同,多足DNA步行器的一个行走步骤同时启动了多个信号的响应性,从而缩短了行走时间并提高了信号放大效率。因此,在最佳条件下,所设计的ECL-RET传感器对HeLa细胞端粒酶活性在1×10至1×10细胞/mL范围内呈现出较宽的动态相关性,检测限低至16细胞/mL。此外,该传感器实现了对不同细胞系中端粒酶活性的通用且可靠的分析。由于在实际样品中具有出色的应用潜力,相信ECL-RET传感系统为端粒酶活性检测在癌症诊断中的应用提供了一种新方法。